Exercise Performance Tests in Mice

Stefan Marcaletti1, Charles Thomas2, Jérôme N. Feige1

1 MusculoSkeletal Diseases, Novartis Institute for Biomedical Research, Basel, Switzerland, 2 Center of Phenogenomics (CPG), Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
Publication Name:  Current Protocols in Mouse Biology
Unit Number:   
DOI:  10.1002/9780470942390.mo100160
Online Posting Date:  March, 2011
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Abstract

Maximal exercise performance is a multifactorial process in which the cardiovascular component, the innervation of the musculature, and the contractile and metabolic properties of skeletal muscle all play key roles. Here, protocols are provided for assessment of maximal running capacity of mice on a treadmill, with a combination of short high‐intensity paradigms primarily intended to test for maximal power and cardiovascular function, and longer low‐intensity paradigms to assess endurance and oxidative metabolism in skeletal muscle. The coupling of treadmill running to indirect calorimetry, to correlate performance measurements to maximal oxygen consumption, is also described. Curr. Protoc. Mouse Biol. 1:141‐154. © 2011 by John Wiley & Sons, Inc.

Keywords: exercise; running; treadmill; VO2max; endurance power

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Measurement of Forced Exercise Performance on a Treadmill
  • Basic Protocol 2: VO2max Determination with a Treadmill Coupled to Indirect Calorimetry
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Measurement of Forced Exercise Performance on a Treadmill

  Materials
  • Appropriate mouse strain (e.g., C57BL/6J) and housing facility
  • Spray bottle of distilled water
  • Mild cleaning agent for cleaning the belt, e.g., 0.75% lysoform
  • Treadmill: see, e.g., Fig. ; the ideal set‐up should include:
    • Revolving belt with adjustable speed (0 to 100 cm/sec)
    • Adjustable slope for up‐ and downhill running (+20°/−20°)
    • Independent lanes with covered tops adapted to the size of mice (∼100 mm width and 450 mm length)
    • A platform at the rear of the belt where the animal can escape when exhausted or in case of a major issue; this platform should, however, be equipped with a system to generate aversive stimulation that forces animals to run during the test (e.g., electrical stimulation grid with adjustable intensity from 0 to 2 mA and automatic detection of each stimulation received)
    • Real‐time control of belt speed, slope, time spent running, distance traveled, and number of aversive stimulations received
    • A number of lanes adapted to the number of animals to assess: commercial systems have been developed, e.g., by Panlab (http://www.panlab.com/), Columbus Instruments (http://www.colinst.com/), and TSE (http://www.tse‐systems.com/)
NOTE: It is recommended but not essential to use a treadmill that communicates with a computer and allows the experimenter to control and record the treadmill parameters (velocity, distance traveled, aversive stimulations per min/cumulative number of aversive stimulations).NOTE: To ensure optimal test results, a few sessions of familiarization with the setup are required a few days preceding the actual treadmill test. For C57BL6/J mice, 1 to 2 sessions is typically sufficient (Fig. ), but this number should be adapted to the particular strain of mice under consideration.

Basic Protocol 2: VO2max Determination with a Treadmill Coupled to Indirect Calorimetry

  Materials
  • Appropriate mouse strain (e.g., C57BL/6J) and housing facility
  • Spray bottle of distilled water
  • Mild cleaning agent for cleaning the belt, e.g., 0.75% lysoform
  • A metabolic treadmill (see, e.g., Fig. ) consisting of a chamber tightly closed at both ends by removable walls (approximate volume, 2 liters); the chamber encompasses:
    • A revolving belt with adjustable speed (0 to 120 cm/sec) adapted to the size of the mice (approximately 50 mm width and 26 cm length)
    • A system at the rear of the belt to encourage animals to run during the test (e.g., electrical stimulation grid with adjustable intensity)
    • An inlet port connected to a pump and an air‐flow controller to ensure chamber ventilation with ambient air at a constant flow
    • An outlet port connected to an air sampler delivering the air inside the chamber to the gas analyzer at regular intervals
    • A fan at the front of the belt to ensure circulation of the air over the animal
    • Ability to adjust the slope of the entire setup in 5° increments from −10° to +25°: commercial systems have been developed, e.g., by Columbus Instruments (http://www.colinst.com/) and TSE (http://www.tse‐systems.com/)
  • An open‐circuit calorimeter (indirect calorimetry) (e.g., Oxymax, Columbus Instruments, http://www.colinst.com/) to monitor changes in gas concentration (O 2 and CO 2) in the air of the chamber; ambient air is used as a reference after calibration of the system with a calibration gas (20.5% O 2/0.5% CO 2 with remainder N 2)
  • Computer and software communicating with the open‐circuit calorimeter to define the settings of the experiment (air flow, sampling time, etc.) and to display measurements in real time (VO 2, VCO 2, and RER) throughout the experiment; ideally, the same software should allow to control and record the treadmill parameters
  • Calibration gas (high‐purity grade): exactly 20.5% O 2 and 0.5% CO 2 with remainder N 2 (custom prepared by gas supplier)
  • Scale to monitor body weight of the mouse
  • Timer
NOTE: To ensure optimal test results, a few sessions of familiarization to the setup are required every day preceding the actual treadmill test. For C57BL6/J mice 1 to 2 sessions is typically sufficient, but this number should be adapted to the particular strain of mice under consideration.
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Figures

Videos

Literature Cited

Literature Cited
   Bernstein, D. 2003. Exercise assessment of transgenic models of human cardiovascular disease. Physiol. Genomics 13:217‐226.
   Billat, V.L., Mouisel, E., Roblot, N., and Melki, J. 2005. Inter‐ and intrastrain variation in mouse critical running speed. J. Appl. Physiol. 98:1258‐1263.
   Booth, F.W., Laye, M.J., and Spangenburg, E.E. 2010. Gold standards for scientists who are conducting animal‐based exercise studies. J. Appl. Physiol. 108:219‐221.
   Dawson, C.A. and Horvath, S.M. 1970. Swimming in small laboratory animals. Med. Sci. Sports 2:51‐78.
   Kemi, O.J., Loennechen, J.P., Wisloff, U., and Ellingsen, O. 2002. Intensity‐controlled treadmill running in mice: Cardiac and skeletal muscle hypertrophy. J. Appl. Physiol. 93:1301‐1309.
   Konhilas, J.P., Maass, A.H., Luckey, S.W., Stauffer, B.L., Olson, E.N., and Leinwand, L.A. 2004. Sex modifies exercise and cardiac adaptation in mice. Am. J. Physiol Heart Circ. Physiol. 287:H2768‐H2776.
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   Lerman, I., Harrison, B.C., Freeman, K., Hewett, T.E., Allen, D.L., Robbins, J., and Leinwand, L.A. 2002. Genetic variability in forced and voluntary endurance exercise performance in seven inbred mouse strains. J. Appl. Physiol. 92:2245‐2255.
   Lightfoot, J.T., Turner, M.J., Debate, K.A., and Kleeberger, S.R. 2001. Interstrain variation in murine aerobic capacity. Med. Sci. Sports Exerc. 33:2053‐2057.
   Peronnet, F. and Aguilaniu, B. 2006. Lactic acid buffering, nonmetabolic CO2 and exercise hyperventilation: A critical reappraisal. Respir. Physiol. Neurobiol. 150:4‐18.
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   Thomas, C., Marcaletti, S., and Feige, J.N. 2011. Assessment of spontaneous locomotor and running activity in mice. Curr. Protoc. Mouse Biol. 1:185‐198.
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